title: Cleavage of native type I collagen by human neutrophil elastase.
abstract: The ability of purified human neutrophil elastase (EC 3.4.21.37) to cleave native type I collagen has been investigated. Soluble human, bovine or rat type I collagen was incubated with neutrophil elastase for 16 h at 25 degrees C before catalysis was stopped with 3, 4-dichloroisocoumarin. Analysis by SDS/PAGE of the collagen digests revealed 3/4-length fragments similar in size to those produced by interstitial collagenase. The collagenolytic activity was dose dependent and was not due to a contaminating metalloproteinase or cysteine proteinase, as it was not inhibited by 1,10-phenanthroline, EDTA or L-trans-epoxysuccinyl-leucylamido-(4-guanidino)butane. The identity of the cleavage products was confirmed using a new antibody that recognizes the unwound alpha2(I)-chain. This detected the 3/4-length fragment of type I collagen following neutrophil elastase cleavage. In addition to cleaving soluble collagen, neutrophil elastase also cleaved reconstituted, radiolabelled type I collagen fibrils, at a rate of 16 microg/min per nmol. These results indicate that neutrophil elastase can cleave native type I collagen in the helix, an activity that might contribute to its roles in connective-tissue pathology.
authors.affiliation: Department of Human Metabolism and Clinical Biochemistry, and Institute for Bone and Joint Medicine, University of Sheffield Medical School, Beech Hill Road, Sheffield S10 2RX, U.K.
authors.forename: W
authors.initials: W
authors.lastname: Kafienah
authors.forename: D J
authors.initials: DJ
authors.lastname: Buttle
authors.forename: D
authors.initials: D
authors.lastname: Burnett
authors.forename: A P
authors.initials: AP
authors.lastname: Hollander
mesh_terms: D000818:Animals; D000906:Antibodies / Q000378:metabolism; D002417:Cattle; D003094:Collagen / Q000378:metabolism*; D006801:Humans; D019272:Leukocyte Elastase / Q000378:metabolism*; D010446:Peptide Fragments / Q000276:immunology; D011480:Protease Inhibitors / Q000494:pharmacology; D011489:Protein Denaturation; D017433:Protein Structure, Secondary; D051381:Rats; D012995:Solubility
chemical_list: D000906:Antibodies; D010446:Peptide Fragments; D011480:Protease Inhibitors; D003094:Collagen; D019272:Leukocyte Elastase